Environmental toxicity studies using chickens as surrogates for wildlife: effects of injection day

Domestic chicken embryos are frequently used for avian developmental toxicity studies of polyhalogenated aromatic hydrocarbons, which are often injected into eggs with oil-based vehicles. Injection times range from immediately prior to incubation (embryonic day zero, E0) to after 4 days of incubation (E4) and beyond. Because the majority of organogenesis in chicken embryos occurs during the first 4 days of development, injection after E0 may miss critical, sensitive, developmental periods. We evaluated whether differences in the day of vehicle administration would lead to differences in standard measures of embryotoxicity. We assessed embryotoxicity using mortality, organ somatic indices, teratogenesis, and behavior in hatchling chickens developmentally exposed to a high volume (1.0 microl/g egg) of corn oil vehicle, which was injected into the airsac at E0 or E4. The E0 vehicle group had 76.5% higher overall embryo mortality, embryos died earlier in development, and hatchlings took more than two times longer to right in a righting reflex test compared to the E4 vehicle group. Other behavioral results demonstrated that hatchling chickens from the E0 vehicle group performed differently from their respective no-inject controls, whereas hatchling chickens from the E4 vehicle group did not. The bursal somatic index differed statistically by injection day and weighed 23.7% more in the E0 vehicle group than the E4 vehicle group. These results suggest that the embryonic day of contaminant injection is an important consideration, particularly when using a high volume of vehicle to evaluate developmental toxicity of a contaminant on embryo mortality or behavior.